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Transcriptional regulation of the neuronal L-type calcium channel α1D subunit gene

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Summary

1. The transcriptional regulation of the rat brain L-type calcium channel α1D subunit (RBα1D) gene was investigated using NG108-15 neuroblastoma-glioma cells.

2. Differentiation of NG108-15 cells in the presence of prostaglandin E1 or retinoic acid resulted in the appearance of mRNA encoding the RBα1D subunit detected using Northern blot analysis.

3. A rat genomic DNA library was screened, and a 15.2-kb clone was isolated and partially sequenced which included part of the 5′ upstream sequence through the initial part of intron 2 of the RBα1D gene.

4. Deletion analysis, using a CAT reporter gene and transfected NG108-15 cells, revealed that the 1.2-kb 5′-upstream sequence from the RBα1D gene contains cis-acting positive and negative regulatory elements. A deletion of the 3′ end of exon 1 also suggested the presence of regulatory elements in the first exon.

5. DNase footprinting of exon 1 of the RBα1D gene revealed two regions protected from digestion by specific protein binding, and the second region included an (ATG)7 trinucleotide repeat sequence. Electrophoretic mobility shift assays confirmed nuclear protein(s) binding to the (ATG)7 sequence.

6. The (ATG)7 sequence functions as a enhancer when linked to a thymidine kinase promoter and a CAT reporter gene.

7. These results provide the initial description of the transcriptional regulation of the RBα1D gene and identify a novel enhancer that consists of an (ATG)7 trinucleotide repeat sequence.

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  1. Michael Mitas

    Present address: Department of Biochemistry and Molecular Biology, Oklahoma State University, 246 Noble Research Center, 74078, Stillwater, Oklahoma

  2. Kay L. Fields

    Present address: ORI, PHS, Room 700, 5515 Security Lane, 20852, Rockville, Maryland

Authors and Affiliations

  1. Division of Cardiology, Department of Medicine, Johns Hopkins University, 720 Rutland Avenue, 21205, Baltimore, Maryland

    Timothy J. Kamp & Eduardo Marban

  2. Laboratory of Biochemical Genetics, National Heart Lung and Blood Institute, NIH, Building 36, Room 1C06, 20892, Bethesda, Maryland

    Michael Mitas, Kay L. Fields, Sadamitsu Asoh, Hemin Chin & Marshall Nirenberg

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  1. Timothy J. Kamp
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Kamp, T.J., Mitas, M., Fields, K.L. et al. Transcriptional regulation of the neuronal L-type calcium channel α1D subunit gene. Cell Mol Neurobiol 15, 307–326 (1995). https://doi.org/10.1007/BF02089942

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